Chinese hamster ovary cells deficient in peroxisomes lack the nonspecific lipid transfer protein (sterol carrier protein 2).
Chinese hamster ovary cells deficient in intact peroxisomes were compared with wild type cells for the presence of the nonspecific lipid transfer protein (nsL-TP; sterol carrier protein 2). With the immunoblotting technique using the affinity purified antibody against rat liver nsL-TP, this protein was shown to be present in the homogenates from wild type cells, but could not be detected in mutant cells. In agreement with a previous study using livers from Zellweger patients it appears that there is a positive, as yet unknown, correlation between peroxisomes and the occurrence of nsL-TP in the cell. As a control using the affinity-purified antibody against the phosphatidylinositol transfer protein from bovine brain, levels of this protein were found to be normal in mutant cells. By use of metrizamide density gradients, nsL-TP was shown to cosediment with a membrane fraction different from peroxisomes. A protein of 58,000 daltons cross-reactive with the antibody against nsL-TP did cosediment with the peroxisomes in wild type cells and possibly with a "peroxisomal remnant" in the mutant cells. Incubation of wild type and mutant cells with L-[3-14C]serine showed that the biosynthesis of phosphatidylserine and the subsequent conversion into phosphatidylethanolamine was comparable in both cell types. This indicates that nsL-TP is not involved in the translocation of phosphatidylserine from the endoplasmic reticulum to the mitochondria as the site of decarboxylation.
van Heusden
G P
GP
Centre for Biomembranes and Lipid Enzymology, State University of Utrecht, The Netherlands.
Bos
K
K
Raetz
C R
CR
Wirtz
K W
KW
D000818:Animals; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics*; D002458:Cell Fractionation; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D015151:Immunoblotting; D008830:Microbodies / Q000648:ultrastructure*; D008970:Molecular Weight; D009154:Mutation; D010053:Ovary; D010716:Phosphatidylinositols / Q000378:metabolism; D010940:Plant Proteins; D013261:Sterols / Q000378:metabolism*
D002352:Carrier Proteins; D010716:Phosphatidylinositols; D010940:Plant Proteins; D013261:Sterols; C072087:sterol carrier proteins